PE Mouse Anti-Human CD51/CD61

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	BM-MSCs and UC-MSCs were harvested by trypsinisation at P0 and P1, pelleted and 20,000 cells per tube were re-suspended in 2% BSA for flow cytometry as described previously. The following antibodies against markers indicative of MSC profiles	Cells were analysed on a FACSCanto II flow cytometer using Diva 7 software (Becton Dickinson & company, Oxford, UK)

Protocol tips
BM-MSCs and UC-MSCs were harvested by trypsinisation at P0 and P1, pelleted and 20,000 cells per tube were re-suspended in 2% BSA for flow cytometry as described previously. The following antibodies against markers indicative of MSC profiles
Downstream tips
Cells were analysed on a FACSCanto II flow cytometer using Diva 7 software (Becton Dickinson & company, Oxford, UK)

Publication protocol

BM-MSCs and UC-MSCs were harvested by trypsinisation at P0 and P1, pelleted and 20,000 cells per tube were re-suspended in 2% BSA for flow cytometry as described previously. The following antibodies against markers indicative of MSC profiles [6] were used: CD90-PE (clone 5E10), CD105-Allophycocyanin (APC) (clone 266), CD73-BV421 (clone AD2), CD19-BV421 (clone HIB19), CD34-APC (clone 581), CD45-PE (clone HI30), HLA-DR-APC (clone TU36) and CD271-BV421 (clone C40-1457). Integrin profiles were also assessed using antibodies against CD29-APC (clone MAR4), CD49a-PE (clone SR84), CD49b-BV421 (clone 12F1), CD49c-PE (clone C3 II.1) and CD51/61-PE (clone 23C6). Chondrogenic potency markers were assessed using antibodies against CD166-BV421 (clone 3A6), CD39-APC (clone TU66), CD44-PerCp-Cy5.5 (clone G44-26) (all from Becton Dickinson & Company, Oxford, UK), CD151-PE (clone 14A2.H1), receptor tyrosine kinase-like orphan receptor 2 (ROR2)-APC (clone 231509) and fibroblastic growth factor receptor3 (FGFR3)-PE (clone 136334) (R&D systems, Abingdon, UK). Immunomodulatory markers were assessed using the following antibodies: CD40-PE (clone 5C3), CD80-PE (clone L307.4), CD86-FITC (clone 2331), CD106-APC (clone 51-10C9) and CD317-PE (clone 26F8). Appropriate isotype-matched IgG controls were used throughout. Cells were analysed on a FACSCanto II flow cytometer using Diva 7 software (Becton Dickinson & company, Oxford, UK)

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